Preparation And Storage
Recommended Assay Procedures
BD™ CompBeads can be used as surrogates to assess fluorescence spillover (Compensation). When fluorochrome conjugated antibodies are bound to CompBeads, they have spectral properties very similar to cells. However, for some fluorochromes there can be small differences in spectral emissions compared to cells, resulting in spillover values that differ when compared to biological controls. It is strongly recommended that when using a reagent for the first time, users compare the spillover on cells and CompBead to ensure that BD Comp beads are appropriate for your specific cellular application.
For optimal and reproducible results, BD Horizon Brilliant Stain Buffer should be used anytime two or more BD Horizon Brilliant dyes are used in the same experiment. Fluorescent dye interactions may cause staining artifacts which may affect data interpretation. The BD Horizon Brilliant Stain Buffer was designed to minimize these interactions. More information can be found in the Technical Data Sheet of the BD Horizon Brilliant Stain Buffer (Cat. No. 563794/566349) or the BD Horizon Brilliant Stain Buffer Plus (Cat. No. 566385).
Product Notices
- The production process underwent stringent testing and validation to assure that it generates a high-quality conjugate with consistent performance and specific binding activity. However, verification testing has not been performed on all conjugate lots.
- An isotype control should be used at the same concentration as the antibody of interest.
- Caution: Sodium azide yields highly toxic hydrazoic acid under acidic conditions. Dilute azide compounds in running water before discarding to avoid accumulation of potentially explosive deposits in plumbing.
- For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
- BD Horizon Brilliant Stain Buffer is covered by one or more of the following US patents: 8,110,673; 8,158,444; 8,575,303; 8,354,239.
- BD Horizon Brilliant Violet 711 is covered by one or more of the following US patents: 8,110,673; 8,158,444; 8,227,187; 8,455,613; 8,575,303; 8,354,239.
- Cy is a trademark of GE Healthcare.
- Alexa Fluor® is a registered trademark of Life Technologies Corporation.
- Please refer to http://regdocs.bd.com to access safety data sheets (SDS).
- Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
Companion Products
The RP-1 monoclonal antibody specifically recognizes the RP-1 Antigen. This cell surface marker is expressed on rat peritoneal and peripheral blood neutrophils. Amongst bone marrow cells, the RP-1 Antigen is expressed on band form and mature neutrophils but is not expressed on promyelocytes, myelocytes, and metamyelocytes. The RP-1 antibody does not bind to either rat monocytes, macrophages, eosinophils or to peritoneal neutrophils from mice, rabbits, guinea pigs, or to human peripheral blood neutrophils. Expression of the RP-1 Antigen on rat peritoneal neutrophils is enhanced by cellular stimulation with Phorbol 12-Myristate 13-Acetate (PMA) or Concanavalin A (ConA). Immunoprecipitation and SDS-PAGE analysis of non-treated and PMA-activated rat neutrophil membranes with the RP-1 antibody revealed two main bands of approximately 85 kDa. The RP-1 antibody is also known as the Mouse Anti-Rat Granulocytes antibody.
The antibody was conjugated to BD Horizon BV711 which is part of the BD Horizon Brilliant™ Violet family of dyes. This dye is a tandem fluorochrome of BD Horizon BV421 with an Ex Max of 405-nm and an acceptor dye with an Em Max at 711-nm. BD Horizon BV711 can be excited by the violet laser and detected in a filter used to detect Cy™5.5 / Alexa Fluor® 700-like dyes (eg, 712/20-nm filter). Due to the excitation and emission characteristics of the acceptor dye, there may be moderate spillover into the Alexa Fluor® 700 and PerCP-Cy5.5 detectors. However, the spillover can be corrected through compensation as with any other dye combination.
Development References (3)
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Gotoh S, Itoh M, Fujii Y, Arai S, Sendo F. Enhancement of the expression of a rat neutrophil-specific cell surface antigen by activation with phorbol myristate acetate and concanavalin A. J Immunol. 1986; 137(2):643-650. (Immunogen: Immunoprecipitation). View Reference
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Kudo C, Araki A, Matsushima K, Sendo F. Inhibition of IL-8-induced W3/25+ (CD4+) T lymphocyte recruitment into subcutaneous tissues of rats by selective depletion of in vivo neutrophils with a monoclonal antibody. J Immunol. 1991; 147(7):2196-2201. (Clone-specific: Immunohistochemistry). View Reference
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Sekiya S, Gotoh S, Yamashita T, Watanabe T, Saitoh S, Sendo F. Selective depletion of rat neutrophils by in vivo administration of a monoclonal antibody. J Leukoc Biol. 1989; 46(2):96-102. (Clone-specific: Immunohistochemistry). View Reference
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Global - Refer to manufacturer's instructions for use and related User Manuals and Technical data sheets before using this products as described
Comparisons, where applicable, are made against older BD Technology, manual methods or are general performance claims. Comparisons are not made against non-BD technologies, unless otherwise noted.
For Research Use Only. Not for use in diagnostic or therapeutic procedures.